New tremella auramtialba strain JSJ-J2HD3-HD25 bred by space breeding technology and application thereof

Abstract

The invention relates to a novel tremella aurantialba strain JSJ-J2HD3-HD25 bred by a space breeding technology and application thereof, and belongs to the technical field of edible mushroom breeding, the novel tremella aurantialba strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.41787, and the preservation number is CGMCC No.41787. The strain and a stereum hirsutum strain JSJ-G2YX001C which is preserved in the China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No.41790 are cultured and bred under a dark culture condition to obtain a yellow tremella aurantialba sporocarp strain. The novel tremella aurantialba strain can omit color changing conditions, save production cost and improve production benefits, can be used as an excellent strain resource for genetic breeding of tremella aurantialba species, researches color-related gene information, and provides an important theoretical basis for gene positioning breeding.

Description

Technical Field

[0001] The present invention belongs to the technical field of edible mushroom breeding, and specifically relates to a new strain of Tremella aurantialba JSJ-J2HD3-HD25 selected by space breeding technology and its application. Background Art

[0002] Tremella aurantialba Naematelia aurantialba belongs to Basidiomycota, Tremellomycetes, Tremellales, Naemateliaceae, and Naematelia Naematelia . The fruiting body of Tremella aurantialba is a heterocomplex formed by Tremella aurantialba and Stereum hirsutum Stereum hirsutum . It has a smooth and tender taste, is rich in nutrients, and contains amino acids, dietary fiber, β-carotene, vitamin B2, etc. The biological characteristics of Tremella aurantialba itself are relatively special, consisting of two species, so the entire breeding system is relatively complex and difficult. At present, most of the newly selected strains of Tremella aurantialba reported publicly are systematic selections, that is, new varieties are selected through domestication and cultivation of wild resources. Systematic selection of new varieties is one of the common methods for variety selection. Although this method is simple to operate and has low cost, its genetic basis range is small, the selection cycle is long, and the efficiency is low. And currently, for the strains of Tremella aurantialba cultivated artificially in the market, the primordia are white under dark culture conditions, and the primordia expand and grow to form fruiting bodies that also remain white. After giving light color conversion conditions during the growth process, the fruiting bodies will turn color to form yellow and then mature. Recently, a report on using space breeding technology to select new varieties of Tremella aurantialba was published (CN119464086A), which disclosed that the total flavonoid content of one strain of Tremella aurantialba fruiting body selected by the Kunming Institute of Botany, Chinese Academy of Sciences through spaceflight was higher than that of Tremella aurantialba without spaceflight.

[0003] Due to the complex biological characteristics of Tremella aurantialba, the development speed of the industry is relatively late compared to other major edible mushrooms, and the development of strain resources is correspondingly less. To ensure the sustainable and healthy development of the Tremella aurantialba industry, it is necessary to have more strains with excellent traits for subsequent gene-directed breeding and other work. Therefore, it is extremely important to develop new varieties of Tremella aurantialba through different breeding methods and screen strains with excellent traits.

[0004] References related to Tremella aurantialba can be found in: [1] Yunnan Mushroom Vision Biotechnology Co., Ltd. A strain of Tremella aurantialba JSJ-J2X1001W and its application and molecular marker identification method: CN118272238A. 2024-07-02. [2] Yunnan Mushroom Vision Biotechnology Co., Ltd. A strain of Tremella aurantialba JSJ-J2F1001C and its application and molecular marker identification method: CN118207101A. 2024-06-18. [3] Yunnan Junshijie Biotechnology Co., Ltd. A Tremella aurantialba strain suitable for industrial cultivation: CN112592835A. April 2, 2021. [4] Kunming Edible Fungi Research Institute, All-China Federation of Supply and Marketing Cooperatives. Two new Tremella aurantialba strains and their SSR molecular marker identification method: CN115261236A. November 1, 2022. [5] Kunming Edible Fungi Research Institute, All-China Federation of Supply and Marketing Cooperatives, Yunnan Supply and Marketing Cooperative Scientific Research Institute. A new Tremella aurantialba strain, Junjin'er No. 1 strain, and its cultivation method: CN111990164A[P]. November 27, 2020. [6] Kunming Institute of Botany, Chinese Academy of Sciences. A space-borne Tremella cinnabarina strain, Hang 8, and its application: CN119464086A. February 18, 2025. [7] Cao Yao, Shen Zhenhui, Yang Linlei, etc. Determination of the polarity of Tremella aurantialba by single spore hybridization technology [J]. Acta Edulis Fungi. 2024, 31(01): 38-44. Summary of the Invention The purpose of the present invention is to provide a new Tremella aurantialba strain JSJ-J2HD3-HD25 selected by space breeding technology and its application.

[0005] The technical solution adopted by the present invention is as follows: A new strain JSJ-J2HD3-HD25 of Tremella aurantialba selected by space breeding technology, and the new strain is deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit number CGMCC No. 41787. Naematelia aurantialba

[0006] Naematelia aurantialba The application of the new strain JSJ-J2HD3-HD25 of Tremella aurantialba selected by space breeding technology in the present invention is to use the new strain JSJ-J2HD3-HD25 to cultivate and produce Tremella aurantialba seeds with the strain JSJ-G2YX001C of Stereum hirsutum. The fruiting bodies of the Tremella aurantialba seeds are yellow from primordium to maturity under dark culture conditions; the Stereum hirsutum strain JSJ-G2YX001C is deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit number CGMCC No. 41790. stereum hirsutum

[0007] ​​​The strain provided by the present invention is yellow from the primordium stage to the mature stage, and does not require additional color-changing conditions to turn yellow. The color-changing conditions can be omitted, saving production costs and improving production efficiency. Moreover, it can be used as an excellent strain resource for the genetic breeding of Tremella aurantialba, study gene information related to color, and provide an important theoretical basis for gene mapping breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 It is a photograph of the biological characteristics of the Tremella aurantialba strain JSJ-J2HD3-HD25; Figure 2 It is a photograph of the biological characteristics of the Stereum hirsutum strain JSJ-G2YX001C; Figure 3 It is the fruiting body of the fungus obtained by culturing the two compatible monokaryotic spore strains D3 and D25 of Tremella aurantialba carried on and Stereum hirsutum JSJ-G2YX001C; Figure 4 It is the fruiting body of the fungus obtained by culturing the two compatible monokaryotic spore strains D3 and D25 of Tremella aurantialba without carrying and Stereum hirsutum JSJ-G2YX001C.

[0009] Figure 5 It is the fruiting body of the fungus obtained by culturing the Tremella aurantialba strain JSJ-J2HD3-HD25 and Stereum hirsutum JSJ-G2YX001C. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0010] The content of the present invention will be further elaborated below in conjunction with specific embodiments.

[0011] A new strain JSJ-J2HD3-HD25 of Tremella aurantialba obtained by space breeding technology, the Tremella aurantialba strain JSJ-J2HD3-HD25 is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is: CGMCC No. 41787, the preservation date is February 19, 2025, and the taxonomic name is Tremella aurantialba Naematelia aurantialba which belongs to the family Naemateliaceae and the genus Naematelia aurantialba Naematelia. It is an asexual spore, the colony is smooth and milky white. See Naematelia ). Figure 1 .

[0012] The said strains were obtained by smearing 44 Tremella aurantialba materials (see Table 1) from Yunnan Junshijie Biotechnology Co., Ltd. on weighing paper. After air-drying the weighing paper, it was placed into 0.2 mL PCR tubes and sent into space for mutagenesis on Shenzhou-15 on November 29, 2022. Among them, there were 10 monokaryotic strains and 34 heterokaryotic strains. The control materials of the above 44 Tremella aurantialba strains were stored in a 4°C refrigerator. All the materials sent into space were returned on July 5, 2023. The materials sent into space and the control materials were simultaneously activated under the culture conditions of PDA medium, 20°C, and darkness. Among them, a total of 64 control and space-sent materials were successfully activated (see Table 1). All the successfully activated control and space-sent materials were cultured and compared with the Stereum hirsutum strain JSJ-G2YX001C stereum hirsutum ), and the fruiting body traits of each control and space-sent material during cultivation were observed for any changes.

[0013] Table 1 List of Tremella aurantialba materials sent into space on Shenzhou-15 by Yunnan Junshijie Biotechnology Co., Ltd. and activation status

[0014]

[0015]

[0016]

[0017] In the table, "√" indicates successful activation; "×" indicates failed activation.

[0018] The Stereum hirsutum strain JSJ-G2YX001C was isolated from the fruiting body of Stereum hirsutum collected in Yuxi City, Yunnan Province. It is preserved in the China General Microbiological Culture Collection Center (CGMCC), with the preservation date of February 19, 2025, and the preservation number: CGMCC No. 41790, and the taxonomic name is Stereum hirsutum stereum hirsutum , which belongs to the family Stereaceae (Stereum hirsutum) and the genus Stereum Stereum ). It is in the mycelial state, with a circular and white colony. See Figure 2 .

[0019] The steps of the culture comparison experiment between the control and space-sent materials and Stereum hirsutum JSJ-G2YX001C are as follows: 1) Prepare the liquid strain of Stereum hirsutum. Medium: 200 g of potatoes (boiled juice), 20 g of glucose, 3 g of potassium dihydrogen phosphate, 1.5 g of magnesium sulfate, 1000 mL of water. After autoclaving and cooling, 20 mycelial agar blocks of strain JSJ-G2YX001C with a size of 5 mm×5 mm×5 mm are inoculated and cultured on a magnetic stirrer at 21 °C with a rotation speed of 180 for 5 days; 2) All successfully activated materials contain Tremella aurantialba monokaryotic spores and asexual spore materials. For the monokaryotic spore materials, 2 compatible strains need to be cultured with Stereum hirsutum JSJ-G2YX001C simultaneously. On PDA medium, 0.5 g of each of the 2 compatible monokaryotic spores successfully cultured for 10 d are picked and mixed with 4 mL of the liquid strain of Stereum hirsutum, and then inoculated into the culture substrate (formula: 30% cottonseed hulls, 56% wood chips, 12% wheat bran, 1% potassium dihydrogen phosphate, 1% calcium sulfate, water content 55% - 65%, pH 6 - 6.5; 130 g of culture substrate is filled in a 240 mL original seed bottle), and cultured at 18 °C, relative humidity 50%, CO2 mass concentration ≤ 800 mg·L -1 , and observe the occurrence of primordia under dark conditions. During this period, transfer the primordia to a new original seed bottle for continuous culture; for the asexual spore materials, directly pick 0.5 g of the spores after the end of culture on PDA medium, mix them with 4 mL of the liquid strain of Stereum hirsutum, and then inoculate them into the culture substrate and culture according to the above conditions; all carried and control materials are cultured and compared according to the above method; 3) A total of 64 strains of all control and carried materials were cultured and cultivated with Stereum hirsutum, and 26 fruiting bodies were obtained. Among them, from the materials carried by spaceflight, Tremella aurantialba fruiting bodies obtained by culturing two compatible Tremella aurantialba monokaryotic spore strains D3 and D25 with Stereum hirsutum JSJ-G2YX001C were yellow from primordium growth to mature fruiting body under dark culture conditions (see Figure 3 ); while the Tremella aurantialba fruiting bodies obtained by culturing the control materials D3 and D25 without spaceflight with Stereum hirsutum JSJ-G2YX001C were white from primordium growth to mature fruiting body under dark conditions (see Figure 4 ), that is, Tremella aurantialba fruiting bodies with a yellow color throughout the growth cycle were obtained by culturing two compatible monokaryotic spore strains D3 and D25 with Stereum hirsutum JSJ-G2YX001C after spaceflight.

[0020] Extract and isolate asexual spores from the yellow Tremella aurantialba fruiting bodies obtained by culturing two compatible monokaryotic spore strains D3 and D25 with Stereum hirsutum JSJ-G2YX001C, and then culture and verify the asexual spores with Stereum hirsutum JSJ-G2YX001C. The relevant extraction, separation, and culture steps are as follows: 1) Harvest the obtained yellow Tremella aurantialba fruit bodies and clean them thoroughly. Wipe the mushroom bodies with 75% alcohol, then rinse with sterile water and place them in a laminar flow hood to air dry. 2) Take 10 pieces of fruit body tissue blocks and place them in a 250 mL liquid medium (the liquid medium is: 200 g of potato juice filtered after boiling, 20 g of glucose, 1000 mL of water); culture under the conditions of a rotation speed of 150 r / min and a temperature of 23 °C for 4 - 5 days; after the culture is completed, transfer 0.5 mL to a PDA medium, culture for 4 - 5 days, then separate the asexual spores in the medium to a new PDA medium and continue to culture for 4 - 5 days. Transfer the asexual spores to a new PDA medium again. After ITS identification, it is identified as Tremella aurantialba, and the asexual spores of Tremella aurantialba are obtained, named JSJ-J2HD3-HD25. 3) Pick 0.5 g of the asexual spores of the JSJ-J2HD3-HD25 strain and mix them with 4 mL of the liquid spawn of Stereum hirsutum JSJ-G2YX001C, and then inoculate them into the culture substrate (formula: 30% cottonseed hulls, 56% wood chips, 12% wheat bran, 1% potassium dihydrogen phosphate, 1% calcium sulfate, water content 55% - 65%, pH 6 - 6.5; 150 g of culture substrate is filled in a 240 mL original seed bottle), and culture at 18 °C, relative humidity 50%, CO2 mass concentration ≤ 800 mg·L -1 , and observe the occurrence of primordia under dark conditions. During this period, transfer the primordia to a new original seed bottle for continuous culture.

[0021] 4) After 40 d of transfer culture through the occurrence of primordia, the asexual spores of the JSJ-J2HD3-HD25 strain and the Stereum hirsutum JSJ-G2YX001C strain also obtained yellow Tremella aurantialba fruit bodies under dark culture conditions, as shown in Figure 5 .

[0022] The growth time of the yellow Tremella aurantialba fruit bodies obtained by culturing the JSJ-J2HD3-HD25 strain was shortened by 5 days compared with the control Tremella aurantialba fruit bodies. It is used to prepare the spawn fruit bodies, and the spawn fruit bodies can be used in about 40 days, while the control spawn fruit bodies need about 45 days to be used for cultivation. The crude polysaccharide nutrient components of the two were detected, indicating that the crude polysaccharide content of the yellow Tremella aurantialba fruit bodies obtained by spaceflight was significantly increased compared with the control Tremella aurantialba fruit bodies (Table 2).

[0023] Table 2 Record table of relevant indexes of spawn fruit bodies cultured with and without spaceflight materials .

Claims

1. Tremella aurantialba strain JSJ-J2HD3-HD25 selected by space breeding technology, characterized in that, Naematelia aurantialba The new strain is deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit number of CGMCC No. 41787. ​ 2. Use of the new strain JSJ-J2HD3-HD25 of Tremella aurantialba selected by space breeding technology as claimed in claim 1, characterized in that, Naematelia aurantialba The new strain JSJ-J2HD3-HD25 is used to cultivate Tremella aurantialba spawn with the strain of Stereum hirsutum ( stereum hirsutum ). The spawn of Tremella aurantialba is a spawn fruiting body that is yellow from primordium to maturity under dark culture conditions. The strain of Stereum hirsutum JSJ-G2YX001C is preserved in the China General Microbiological Culture Collection Center with the preservation number of CGMCC No. 41790.

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